Farah Rauf Shakoori

Isolation and characterization of arsenic resistant bacteria from wastewater

The present study proposed the isolation of arsenic resistant bacteria from wastewater. Only three bacterial isolates (MNZ1, MNZ4 and MNZ6) were able to grow in high concentrations of arsenic. The minimum inhibitory concentrations of arsenic against MNZ1, MNZ4 and MNZ6 were 300 mg/L, 300 mg/L and 370 mg/L respectively. The isolated strains showed maximum growth at 37 °C and at 7.0 pH in control but in arsenite stress Luria Bertani broth the bacterial growth is lower than control. All strains were arsenite oxidizing. All strains were biochemically characterized and ribotyping (16S rRNA) was done for the purpose of identification which confirmed that MNZ1 was homologous to Enterobacter sp. while MNZ4 and MNZ6 showed their maximum homology with Klebsiella pneumoniae. The protein profiling of these strains showed in arsenic stressed and non stressed conditions, so no bands of induced proteins appeared in stressed conditions. The bacterial isolates can be exploited for bioremediation of arsenic containing wastes, since they seem to have the potential to oxidize the arsenite (more toxic) into arsenate (less toxic) form.

Electrochemistry and microbiology of microbial fuel cells treating marine sediments polluted with heavy metals

The industrial contamination of marine sediments with chromium, copper and nickel in Penang, Malaysia was addressed with bio-remediation, coupled with power generation, using in situ sediment microbial cells (SMFCs) under various conditions. The efficiency of aerated sediment microbial fuel cells (A-SMFCs) and non-aerated sediment microbial fuel cells (NA-SMFCs) was studied. The A-SMFCs generated a voltage of 580.5 mV between 50 and 60 days, while NA-SMFCs produced a voltage of 510 mV between 60 and 80 days. The cell design point for A-SMFCs was 2 kΩ, while for NA-SMFCs it was 200 Ω. In both SMFCs, the maximum current values relating to forward scanning, reverse scanning and oxidation/reduction peaks were recorded on the 80th day. The anode showed maximum additional capacitance on the 80th day (A-SMFC: 2.7 F cm−2; and NA-SMFC: 2.2 F cm−2). The whole cell electrochemical impedance using the Nyquist model was 21 Ω for A-SMFCs and 15 Ω for NA-SMFCs. After glucose enrichment, the impedance of A-SMFCs was 24.3 Ω and 14.6 Ω for NA-SMFCs. After 60 days, the A-SMFCs reduced the maximum amount of Cr(VI) to Cr(III) ions (80.70%) and Cu(II) to Cu(I) ions (72.72%), and showed maximum intracellular uptake of Ni(II) ions (80.37%); the optimum remediation efficiency of NA-SMFCs was after 80 days toward Cr(VI) ions (67.36%), Cu(II) ions (59.36%) and Ni(II) ions (52.74%). Both SMFCs showed highest heavy metal reduction and power generation at a pH of 7.0. SEM images and 16S rRNA gene analysis showed a diverse bacterial community in both A-SMFCs and NA-SMFCs. The performance of A-SMFCs showed that they could be exercised as durable and efficient technology for power production and the detoxification of heavy metal sediments. The NA-SMFCs could also be employed where anaerobic fermentation is required.

Molecular Characterization of a Copper Metallothionein Gene from a Ciliate Tetrahymena farahensis

A new copper metallothionein (TfCuMT) gene has been identified from a locally isolated ciliate Tetrahymena farahensis. It contains 327 nucleotides encoding a peptide chain of 108 amino acids and belongs to class MTT2 and subfamily 7b. Amplification from both gDNA and mRNA confirmed the intronless nature of this gene. Like most of the metallohtioneins, cysteine residues contribute nearly 30% content with the specific CKC motifs. Structural repeats present in peptide sequence of TfCuMT indicate internal duplication of gene at some stage of gene evolution. The predicted model of copper metallothionein protein showed that copper ions are mainly chelated by thiol sulfur of cysteine residues and are embedded in the folds of polypeptide chain. For in vivo expression of TfCuMT in Escherichia coli host cells the classical stop codons, which coded for glutamine in the ciliate were mutated to CAA and CAG through site directed mutagenesis. The mutated gene showed higher expression in pET28a expression vector compared with pET21a. Optimum expression was obtained after 6–8 h of 0.1 mM IPTG induction. Stability of His tagged TfCuMT in 5% SDS was low, with half‐life of about 104 min. Presence of 1.0 μM copper increased the expression level by 1.65‐fold. Presence of 100 μM Cysteine in culture medium caused 2.4‐fold increase in expression level. His tagged TfCuMT was purified through affinity chromatography using NTN‐His binding resin in the presence of 0.1 M imidazole and NaCl. The modeled structure of the TfCuMT showed a cleft for Cu binding with correct orientation of Cys residues in the motif CKC. J. Cell. Biochem. 117: 1843–1854, 2016.

Arsenic processing of yeast isolates IIB-As1 & IIB-As2 and production of glutathione under stress conditions

Four arsenic resistant yeast were isolated from the industrial wastewater. Two strains IIB‐As1 and IIB‐As2 identified as Candida tropicalis and Saccharomyces cerevisiae, respectively. IIB‐As1 and IIB‐As2 showed maximum arsenic resistance. IIB‐As1 showed maximum growth at 35 °C whereas it was 30 °C for IIB‐As2. The yeast isolate showed typical growth curves, but arsenic extended the lag phase. Glutathione plays an important role in metal tolerance. In the present study, As increased the level glutathione and non‐protein thiols in yeast isolates. Removal of As from supernatant was analyzed using the atomic absorption spectrophotometer. They removed arsenic from the medium after 72 h of incubation. Both yeast strains efficiently removed arsenic from the industrial effluent when used individually or in consortia.

Variability in Secondary Structure of 18S Ribosomal RNA as Topological Marker for Identification of Paramecium species

Besides cytological and molecular applications, Paramecium is being used in water quality assessment and for determination of saprobic levels. An unambiguous identification of these unicellular eukaryotes is not only essential, but its ecological diversity must also be explored in the local environment. 18SrRNA genes of all the strains of Paramecium species isolated from waste water were amplified, cloned and sequenced. Phylogenetic comparison of the nucleotide sequences of these strains with 23 closely related Paramecium species from GenBank Database enabled identification of Paramecium multimicronucleatum and Paramecium jenningsi. Some isolates did not show significant close association with other Paramecium species, and because of their unique position in the phylogenetic tree, they were considered new to the field. In the present report, these isolates are being designated as Paramecium caudatum pakistanicus. In this article, secondary structure of 18SrRNA has also been analyzed as an additional and perhaps more reliable topological marker for species discrimination and for determining possible phylogenetic relationship between the ciliate species. On the basis of comparison of secondary structure of 18SrRNA of various isolated Paramacium strains, and among Paramecium caudatum pakistanicus, Tetrahymena thermophila, Drosophila melanogaster, and Homo sapiens, it can be deduced that variable regions are more helpful in differentiating the species at interspecific level rather than at intraspecific level. It was concluded that V3 was the least variable region in all the organisms, V2 and V7 were the longest expansion segments of D. melanogaster and there was continuous mutational bias towards G.C base pairing in H. sapiens. J. Cell. Biochem. 115: 2077–2088, 2014.

Numerical Changes in Chromosomes

Every organism has basic specific number of chromosomes, which are constant for a species. Changes in the chromosomal number, however, do occur which reflect high inviability and phenotypic anomalies. Abnormal euploidy will result if whole set of chromosome is involved, and aneuploidy will result if parts of chromosomal set are involved. The most common abnormal euploids are polyploids such as triploids, tetraploids, etc. Allopolyploids can be made by crossing and doubling progeny chromosomes with colchicines. This technique has important applications in crop breeding. Aneuploids result in an unbalanced genotype with an abnormal phenotype. Monosomy (2n−1) and trisomy (2n + 1) are examples of aneuploid. Aneuploid conditions such as Down syndrome, Klinefelter syndrome, and Turner syndrome are well studied in humans. It is believed to be due to chromosomal nondisjunction and constitutes major portion of genetically based ill-health in human population.